Bulk B Cell analysis via NGS is a method used to explore the repertoire of B cell receptors (BCRs) in a sample that contains heterogeneous B cell populations. This technique provides a comprehensive view of the dynamic changes that occur in the B cell populations during the development of the immune response. Researchers use this technique to analyze the immune repertoire during vaccination experiments, immune infections, and autoimmune disease research.
Bulk B Cell analysis via NGS begins with the extraction of RNA from the sample containing B cells. The RNA is then reverse transcribed into cDNA and subsequently amplified using primers that target the variable regions of the immunoglobulin heavy and light chains. Following PCR amplification, amplicons are sequenced using high-throughput sequencing.
Once the dataset has been generated, bioinformaticians process the data to filter out poor sequences, exclude sequencing artifacts, and extract cell-specific BCR sequences. These sequences are then clustered into B cell clonotypes, which represent individual clones of B cells that share common CDR3 sequences. The resulting data provides information on each clone's size, frequency, and diversity within the population. Visualization tools like heatmaps, PCA plots, and other analytical tools are used to analyze the B cell population diversity and clonotype overlap to better understand the immune response.